The development of Lactococcus lactis as an antimicrobial agent

Non-pathogenic lactic acid bacteria are economically important Gram-positive bacteria used extensively in the food industry. Due to their “generally regarded as safe” status, certain species from the genera Lactobacillus and Lactococcus are also considered desirable as candidates for the production and secretion of recombinant proteins, particular those with therapeutic applications. The hypothesis examined by this thesis is that Lactococcus lactis can be modified to be an effective antimicrobial agent. Therefore, the aims of this thesis were to investigate the optimisation of the expression, secretion and/or activities of potential heterologous antimicrobial proteins by the model lactic acid bacterium, Lactococcus lactis subsp. cremoris MG1363. L. lactis strains were engineered to express and secrete the recombinant CyuC surface protein from Lactobacillus reuteri BR11, and a fusion protein consisting of CyuC and lysostaphin using the Sep promoter and secretion signal. CyuC has been characterised as a cystine-binding protein, but has also been demonstrated to have fibronectin binding activity. Lysostaphin is a bacteriolytic enzyme with specific activity against the Gram-positive pathogen, Staphylococcus aureus. These modified L. lactis strains were then investigated to see if they had the ability to inhibit the adhesion of S. aureus to host extracellular matrix (ECM) proteins. It was observed that the cell extracts of the L. lactis strain with the vector only (pGhost9:ISS1) was able to inhibit the adhesion of S. aureus to fibronectin, whilst the cell extracts of the L. lactis strain expressing lysostaphin was able to inhibit adhesion to keratin. Finally, this thesis has identified specific lactococcal genes that affect the secretion of lysostaphin through the use of random transposon mutagenesis. Ten mutants with higher lysostaphin activity contained insertions in four different genes encoding: (i) an uncharacterised putative transmembrane protein (llmg_0609), (ii) an enzyme catalysing the first step in peptidoglycan biosynthesis (murA2), (iii) a homolog of the oxidative defence regulator (trmA), and (iv) an uncharacterised putative enzyme involved in ubiquinone biosynthesis (llmg_2148). The higher lysostaphin activity observed in these mutants was found to be due to higher amounts of lysostaphin being secreted. The findings of this thesis contribute to the development of this organism as an antimicrobial agent and also to our understanding of L. lactis genetics.

Formalin fixation affects equilibrium partitioning of an ionic contrast agent-microcomputed tomography (EPIC-[mu]CT) imaging of osteochondral samples

Equilibrium Partitioning of an Ionic Contrast agent with microcomputed tomography (EPIC-[mu]CT) is a non-invasive technique to quantify and visualize the three-dimensional distribution of glycosaminoglycans (GAGs) in fresh cartilage tissue. However, it is unclear whether this technique is applicable to already fixed tissues. Therefore, this study aimed at investigating whether formalin fixation of bovine cartilage affects X-ray attenuation, and thus the interpretation of EPIC-[mu]CT data.Design Osteochondral samples (n = 24) were incubated with ioxaglate, an ionic contrast agent, for 22 h prior to [mu]CT scanning. The samples were scanned in both formalin-fixed and fresh conditions. GAG content was measured using a biochemical assay and normalized to wet weight, dry weight, and water content to determine potential reasons for differences in X-ray attenuation.Results The expected zonal distribution of contrast agent/GAGs was observed for both fixed and fresh cartilage specimens. However, despite no significant differences in GAG concentrations or physical properties between fixed and fresh samples, the average attenuation levels of formalin-fixed cartilage were 14.3% lower than in fresh samples.Conclusions EPIC-[mu]CT is useful for three-dimensional visualization of GAGs in formalin-fixed cartilage. However, a significant reduction in X-ray attenuation for fixed (compared to fresh) cartilage must be taken into account and adjusted for accordingly when quantifying GAG concentrations using EPIC-[mu]CT.

Internal market orientation and market-oriented behaviours

Purpose: The purpose of this empirical paper is to investigate internal marketing from a behavioural perspective. The impact of internal marketing behaviours, operationalised as an internal market orientation (IMO), on employees’ marketing and other in-role behaviours (IRB) were examined. ---------- Design/methodology/approach: Survey data measuring IMO, market orientation and a range of constructs relevant to the nomological network in which they are embedded were collected from the UK retail managers. These were tested to establish their psychometric properties and the conceptual model was analysed using structural equations modelling, employing a partial least squares methodology. ---------- Findings: IMO has positive consequences for employees’ market-oriented and other IRB. These, in turn, influence marketing success. Research limitations/implications – The paper provides empirical support for the long-held assumption that internal and external marketing are related and that organisations should balance their external focus with some attention to employees. Future research could measure the attitudes and behaviours of managers, employees and customers directly and explore the relationships between them. ---------- Practical implications: Firm must ensure that they do not put the needs of their employees second to those of managers and shareholders; managers must develop their listening skills and organisations must become more responsive to the needs of their employees. ---------- Originality/value: The paper contributes to the scarce body of empirical support for the role of internal marketing in services organisations. For researchers, this paper legitimises the study of internal marketing as a route to external market success; for managers, the study provides quantifiable evidence that focusing on employees’ wants and needs impacts their behaviours towards the market.

The Spartan-3E Tutorial 1 : Introduction to FPGA Programming, Version 1.0

This tutorial is designed to help new users become familiar with using the Spartan-3E board. The tutorial steps through the following: writing a small program in VHDL which carries out simple combinational logic; connecting the program inputs and outputs to the switches, buttons and LEDs on the Spartan-3E board; and downloading the program to the Spartan-3E board using the Project Navigator software.

Engaging operations management students by designing the unit with industry focus

Operations management is an area concerned with the production of goods and services ensuring that business operations are efficient in utilizing resource and effective to meet customer requirements. It deals with the design and management of products, processes, services and supply chains and considers the acquisition, development, and effective and efficient utilization of resources. Unlike other engineering subjects, content of these units could be very wide and vast. It is therefore necessary to cover the content that is most related to the contemporary industries. It is also necessary to understand what engineering management skills are critical for engineers working in the contemporary organisations. Most of the operations management books contain traditional Operations Management techniques. For example ‘inventory management’ is an important topic in operations management. All OM books deal with effective method of inventory management. However, new trend in OM is Just in time (JIT) delivery or minimization of inventory. It is therefore important to decide whether to emphasise on keeping inventory (as suggested by most books) or minimization of inventory. Similarly, for OM decisions like forecasting, optimization and linear programming most organisations now a day’s use software. Now it is important for us to determine whether some of these software need to be introduced in tutorial/ lab classes. If so, what software? It is established in the Teaching and Learning literature that there must be a strong alignment between unit objectives, assessment and learning activities to engage students in learning. Literature also established that engaging students is vital for learning. However, engineering units (more specifically Operations management) is quite different from other majors. Only alignment between objectives, assessment and learning activities cannot guarantee student engagement. Unit content must be practical oriented and skills to be developed should be those demanded by the industry. Present active learning research, using a multi-method research approach, redesigned the operations management content based on latest developments in Engineering Management area and the necessity of Australian industries. The redesigned unit has significantly helped better student engagement and better learning. It was found that students are engaged in the learning if they find the contents are helpful in developing skills that are necessary in their practical life.

QoS-oriented resource allocation and scheduling of multiple composite Web services in a hybrid cloud using a random-key genetic algorithm

In cloud computing resource allocation and scheduling of multiple composite web services is an important challenge. This is especially so in a hybrid cloud where there may be some free resources available from private clouds but some fee-paying resources from public clouds. Meeting this challenge involves two classical computational problems. One is assigning resources to each of the tasks in the composite web service. The other is scheduling the allocated resources when each resource may be used by more than one task and may be needed at different points of time. In addition, we must consider Quality-of-Service issues, such as execution time and running costs. Existing approaches to resource allocation and scheduling in public clouds and grid computing are not applicable to this new problem. This paper presents a random-key genetic algorithm that solves new resource allocation and scheduling problem. Experimental results demonstrate the effectiveness and scalability of the algorithm.

Design of a model-generated repository as a service for USDL

SAP and its research partners have been developing a lan- guage for describing details of Services from various view- points called the Unified Service Description Language (USDL). At the time of writing, version 3.0 describes technical implementation aspects of services, as well as stakeholders, pricing, lifecycle, and availability. Work is also underway to address other business and legal aspects of services. This language is designed to be used in service portfolio management, with a repository of service descriptions being available to various stakeholders in an organisation to allow for service prioritisation, development, deployment and lifecycle management. The structure of the USDL metadata is specified using an object-oriented metamodel that conforms to UML, MOF and EMF Ecore. As such it is amenable to code gener-ation for implementations of repositories that store service description instances. Although Web services toolkits can be used to make these programming language objects available as a set of Web services, the practicalities of writing dis- tributed clients against over one hundred class definitions, containing several hundred attributes, will make for very large WSDL interfaces and highly inefficient “chatty” implementations. This paper gives the high-level design for a completely model-generated repository for any version of USDL (or any other data-only metamodel), which uses the Eclipse Modelling Framework’s Java code generation, along with several open source plugins to create a robust, transactional repository running in a Java application with a relational datastore. However, the repository exposes a generated WSDL interface at a coarse granularity, suitable for distributed client code and user-interface creation. It uses heuristics to drive code generation to bridge between the Web service and EMF granularities.

Tangible programming elements for young children

Tangible programming elements offer the dynamic and programmable properties of a computer without the complexity introduced by the keyboard, mouse and screen. This paper explores the extent to which programming skills are used by children during interactions with a set of tangible programming elements: the Electronic Blocks. An evaluation of the Electronic Blocks indicates that children become heavily engaged with the blocks, and learn simple programming with a minimum of adult support.

Unpacking Japan's 21st century "National Conversation" : images of the future beyond the iron cage of the "Catch Up" model

How does the image of the future operate upon history, and upon national and individual identities? To what extent are possible futures colonized by the image? What are the un-said futurecratic discourses that underlie the image of the future? Such questions inspired the examination of Japan’s futures images in this thesis. The theoretical point of departure for this examination is Polak’s (1973) seminal research into the theory of the ‘image of the future’ and seven contemporary Japanese texts which offer various alternative images for Japan’s futures, selected as representative of a ‘national conversation’ about the futures of that nation. These seven images of the future are: 1. Report of the Prime Minister’s Commission on Japan’s Goals in the 21st Century—The Frontier Within: Individual Empowerment and Better Governance in the New Millennium, compiled by a committee headed by Japan’s preeminent Jungian psychologist Kawai Hayao (1928-2007); 2. Slow Is Beautiful—a publication by Tsuji Shinichi, in which he re-images Japan as a culture represented by the metaphor of the sloth, concerned with slow and quality-oriented livingry as a preferred image of the future to Japan’s current post-bubble cult of speed and economic efficiency; 3. MuRatopia is an image of the future in the form of a microcosmic prototype community and on-going project based on the historically significant island of Awaji, and established by Japanese economist and futures thinker Yamaguchi Kaoru; 4. F.U.C.K, I Love Japan, by author Tanja Yujiro provides this seven text image of the future line-up with a youth oriented sub-culture perspective on that nation’s futures; 5. IMAGINATION / CREATION—a compilation of round table discussions about Japan’s futures seen from the point of view of Japan’s creative vanguard; 6. Visionary People in a Visionless Country: 21 Earth Connecting Human Stories is a collection of twenty one essays compiled by Denmark born Tokyo resident Peter David Pedersen; and, 7. EXODUS to the Land of Hope, authored by Murakami Ryu, one of Japan’s most prolific and influential writers, this novel suggests a future scenario portraying a massive exodus of Japan’s youth, who, literate with state-of-the-art information and communication technologies (ICTs) move en masse to Japan’s northern island of Hokkaido to launch a cyber-revolution from the peripheries. The thesis employs a Futures Triangle Analysis (FTA) as the macro organizing framework and as such examines both pushes of the present and weights from the past before moving to focus on the pulls to the future represented by the seven texts mentioned above. Inayatullah’s (1999) Causal Layered Analysis (CLA) is the analytical framework used in examining the texts. Poststructuralist concepts derived primarily from the work of Michel Foucault are a particular (but not exclusive) reference point for the analytical approach it encompasses. The research questions which reflect the triangulated analytic matrix are: 1. What are the pushes—in terms of current trends—that are affecting Japan’s futures? 2. What are the historical and cultural weights that influence Japan’s futures? 3. What are the emerging transformative Japanese images of the future discourses, as embodied in actual texts, and what potential do they offer for transformative change in Japan? Research questions one and two are discussed in Chapter five and research question three is discussed in Chapter six. The first two research questions should be considered preliminary. The weights outlined in Chapter five indicate that the forces working against change in Japan are formidable, structurally deep-rooted, wide-spread, and under-recognized as change-adverse. Findings and analyses of the push dimension reveal strong forces towards a potentially very different type of Japan. However it is the seven contemporary Japanese images of the future, from which there is hope for transformative potential, which form the analytical heart of the thesis. In analyzing these texts the thesis establishes the richness of Japan’s images of the future and, as such, demonstrates the robustness of Japan’s stance vis-à-vis the problem of a perceived map-less and model-less future for Japan. Frontier is a useful image of the future, whose hybrid textuality, consisting of government, business, academia, and creative minority perspectives, demonstrates the earnestness of Japan’s leaders in favour of the creation of innovative futures for that nation. Slow is powerful in its aim to reconceptualize Japan’s philosophies of temporality, and build a new kind of nation founded on the principles of a human-oriented and expanded vision of economy based around the core metaphor of slowness culture. However its viability in Japan, with its post-Meiji historical pushes to an increasingly speed-obsessed social construction of reality, could render it impotent. MuRatopia is compelling in its creative hybridity indicative of an advanced IT society, set in a modern day utopian space based upon principles of a high communicative social paradigm, and sustainability. IMAGINATION / CREATION is less the plan than the platform for a new discussion on Japan’s transformation from an econo-centric social framework to a new Creative Age. It accords with emerging discourses from the Creative Industries, which would re-conceive of Japan as a leading maker of meaning, rather than as the so-called guzu, a term referred to in the book meaning ‘laggard’. In total, Love Japan is still the most idiosyncratic of all the images of the future discussed. Its communication style, which appeals to Japan’s youth cohort, establishes it as a potentially formidable change agent in a competitive market of futures images. Visionary People is a compelling image for its revolutionary and subversive stance against Japan’s vision-less political leadership, showing that it is the people, not the futures-making elite or aristocracy who must take the lead and create a new vanguard for the nation. Finally, Murakami’s Exodus cannot be ruled out as a compelling image of the future. Sharing the appeal of Tanja’s Love Japan to an increasingly disenfranchised youth, Exodus portrays a near-term future that is achievable in the here and now, by Japan’s teenagers, using information and communications technologies (ICTs) to subvert leadership, and create utopianist communities based on alternative social principles. The principal contribution from this investigation in terms of theory belongs to that of developing the Japanese image of the future. In this respect, the literature reviews represent a significant compilation, specifically about Japanese futures thinking, the Japanese image of the future, and the Japanese utopia. Though not exhaustive, this compilation will hopefully serve as a useful starting point for future research, not only for the Japanese image of the future, but also for all image of the future research. Many of the sources are in Japanese and their English summations are an added reason to respect this achievement. Secondly, the seven images of the future analysed in Chapter six represent the first time that Japanese image of the future texts have been systematically organized and analysed. Their translation from Japanese to English can be claimed as a significant secondary contribution. What is more, they have been analysed according to current futures methodologies that reveal a layeredness, depth, and overall richness existing in Japanese futures images. Revealing this image-richness has been one of the most significant findings of this investigation, suggesting that there is fertile research to be found from this still under-explored field, whose implications go beyond domestic Japanese concerns, and may offer fertile material for futures thinkers and researchers, Japanologists, social planners, and policy makers.

A hierarchical security assessment model for object-oriented programs

We present a hierarchical model for assessing an object-oriented program's security. Security is quantified using structural properties of the program code to identify the ways in which `classified' data values may be transferred between objects. The model begins with a set of low-level security metrics based on traditional design characteristics of object-oriented classes, such as data encapsulation, cohesion and coupling. These metrics are then used to characterise higher-level properties concerning the overall readability and writability of classified data throughout the program. In turn, these metrics are then mapped to well-known security design principles such as `assigning the least privilege' and `reducing the size of the attack surface'. Finally, the entire program's security is summarised as a single security index value. These metrics allow different versions of the same program, or different programs intended to perform the same task, to be compared for their relative security at a number of different abstraction levels. The model is validated via an experiment involving five open source Java programs, using a static analysis tool we have developed to automatically extract the security metrics from compiled Java bytecode.

Train schedule coordination at an interchange station through Agent negotiation

In open railway markets, coordinating train schedules at an interchange station requires negotiation between two independent train operating companies to resolve their operational conflicts. This paper models the stakeholders as software agents and proposes an agent negotiation model to study their interaction. Three negotiation strategies have been devised to represent the possible objectives of the stakeholders, and they determine the behavior in proposing offers to the proponent. Empirical simulation results confirm that the use of the proposed negotiation strategies lead to outcomes that are consistent with the objectives of the stakeholders.

Structural reliability prediction of a steel bridge element using Dynamic Object Oriented Bayesian Network (DOOBN)

Different from conventional methods for structural reliability evaluation, such as, first/second-order reliability methods (FORM/SORM) or Monte Carlo simulation based on corresponding limit state functions, a novel approach based on dynamic objective oriented Bayesian network (DOOBN) for prediction of structural reliability of a steel bridge element has been proposed in this paper. The DOOBN approach can effectively model the deterioration processes of a steel bridge element and predict their structural reliability over time. This approach is also able to achieve Bayesian updating with observed information from measurements, monitoring and visual inspection. Moreover, the computational capacity embedded in the approach can be used to facilitate integrated management and maintenance optimization in a bridge system. A steel bridge girder is used to validate the proposed approach. The predicted results are compared with those evaluated by FORM method.

Queensland requirements (old and new) for the appointment of a real estate agent

In Moneywood Pty Ltd v Salamon Nominees Pty Ltd 1 the High Court of Australia considered an appeal from the Queensland Court of Appeal in relation to the correct interpretation of s76 (1)(c) Auctioneers and Agents Act 1971 (Qld). In paraphrase, s76(1)(c) provides that a real estate agent shall not be entitled to sue for or recover any commission unless “the engagement or appointment to act as …..real estate agent ….. in respect of such transaction is in writing signed by the person to be charged with such…..commission…..or the person’s agent or representative” (“the statutory requirement”).

The spermostatic and microbicidal actions of quinones and malemides : towards a dual-purpose contraceptive agent

There is an urgent need to develop safe, effective, dual-purpose contraceptive agents that combine the prevention of pregnancy with protection against sexually transmitted diseases. Here we report the identification of a group of compounds that on contact with human spermatozoa induce a state of “spermostasis,” characterized by the extremely rapid inhibition of sperm movement without compromising cell viability. These spermostatic agents were more active and significantly less toxic than the reagent in current clinical use, nonoxynol 9, giving therapeutic indices (ratio of spermostatic to cytotoxic activity) that were orders of magnitude greater than this traditional spermicide. Although certain compounds could trigger reactive oxygen species generation by spermatozoa, this activity was not correlated with spermostasis. Rather, the latter was associated with alkylation of two major sperm tail proteins that were identified as A Kinase-Anchoring Proteins (AKAP3 and AKAP4) by mass spectrometry. As a consequence of disrupted AKAP function, the abilities of cAMP to drive protein kinase A-dependent activities in the sperm tail, such as the activation of SRC and the consequent stimulation of tyrosine phosphorylation, were suppressed. Furthermore, analysis of microbicidal activity using Chlamydia muridarum revealed powerful inhibitory effects at the same low micromolar doses that suppressed sperm movement. In this case, the microbicidal action was associated with alkylation of Major Outer Membrane Protein (MOMP), a major chlamydial membrane protein. Taken together, these results have identified for the first time a novel set of cellular targets and chemical principles capable of providing simultaneous defense against both fertility and the spread of sexually transmitted disease.